Purification and Characterization of Recombinant Human Interleukin 5 Expressed in Chinese Hamster Ovary Cells

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Abstract
Recombinant human interleukin 5 (rhIL-5) expressed in Chinese hamster ovary cells was purified and characterized. Molecular heterogeneity was observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Two major components of Mr around 40, 000 were detected under non-reducing conditions. However, under reducing conditions, the Mr of rhIL-5 was determined to be 22, 000 and 20, 000. After treatment with endoglycosidase F, a band with an apparent MT of 18, 000 was observed. Treatment of rhIL-5 with 2-mercaptoethanol followed by N-ethylmaleimide resulted in its dissociation into a mono-meric form. This alkylated rhIL-5 was biologically less active than intact rhIL-5. These results suggest that rhIL-5 exists as a dimer, and that the heterogeneity of rhIL-5 is mainly due to the difference in the content of carbohydrate. Moreover, the formation of disulfide bond(s) might be important for the biological activity of rhIL-5.

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